Baicalin,a flavonoid,affects the activity of human dermal papilla cells and promotes anagen induction in mice

Baicalin, a flavonoid isolated from Scutellaria baicalensis, is known to have multiple biological functions. Recent studies have demonstrated that baicalin treatment increases alkaline phosphatase activity (ALP) and osteoprotegerin secretion by osteoblasts. Furthermore, baicalin induces the differentiation of cultured osteoblasts via the activation of the Wnt/β-catenin signaling pathway. In this study, we evaluated the hair growth-promoting effects of baicalin in human follicular dermal papilla (DP) cells. A reporter assay and Western blotting were used to assess the effect of baicalin on β-catenin signaling in DP cells. ALP activity and messenger RNA (mRNA) expression were examined by ALP staining and real-time polymerase chain reaction (PCR), respectively. Growth factor expression levels were also evaluated using real-time PCR. Finally, the effect of baicalin on hair growth in vivo was examined by topical application of baicalin on the shaved dorsal skin of C57BL/6 mice. Our results indicate that baicalin activates Wnt/β-catenin signaling in a dose-dependent manner in human DP cells. ALP mRNA expression and activity were significantly induced in the presence of baicalin. In addition, treatment with baicalin induced the mRNA expression of growth factors, such as insulin-like growth factor-1 (IGF-1) and vascular endothelial growth factor (VEGF). Moreover, compared to vehicle treatment, baicalin treatment induced an earlier conversion from telogen to anagen. Our results strongly suggest that baicalin promotes hair growth by regulating the activity of DP cells.

     

Hair growth regulation by the extract of aromatic plant <Emphasis Type="Italic">Erica multiflora</Emphasis>

Hair growth problems can affect human physical and mental health and are of particular relevance during the aging process. In an effort to resolve such problems, we attempted to find plants having hair growth regulation activity and thus collected plant extracts from Tunisia for bioprospecting purposes. Among them, we investigated the Erica multiflora extract to evaluate the hair growth promotion activity by using the MTT (3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide) assay and cell cycle assay on human dermal papilla cells in vitro and an administration assay on mouse dorsal skin in vivo. The results showed that the Erica multiflora extract promotes dermal papilla cell growth and cell cycle with high activity, and induced hair growth in vivo by induction of anagen phase from telogen phase.     

N-benzyl-5-phenyl-1H-pyrazole-3-carboxamide promotes vascular endothelial cell angiogenesis and migration in the absence of serum and FGF-2

Aim:

To investigate the effect of N-benzyl-5-phenyl-1H-pyrazole-3-carboxamide (BPC) on angiogenesis in human umbilical vein endothelial cells (HUVECs).

Methods:

Capillary-like tube formation on matrigel and cell migration analyses were performed in the absence of serum and fibroblast growth factor (FGF-2). Reactive oxygen species (ROS) were measured using a fluorescent probe, 2′, 7′- dichlorodihydrofluorescein (DCHF). The nitric oxide (NO) production of HUVECs was examined using a NO detection kit. Morphological observation under a phase contrast microscope, a viability assay using 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyl-tetrazolium (MTT) and a lactate dehydrogenase (LDH) activity analysis by a detection kit were performed to evaluate the toxicity of BPC on HUVECs in the presence of serum and FGF-2. The level of hypoxia-inducible factor 1α (HIF-1α) and the release of vascular endothelial growth factor (VEGF) were measured by Western blot and ELISA, respectively.

Results:

In the absence of serum and FGF-2, cells treated with BPC (5-20 μmol/L) rapidly aligned with one another and formed tube-like structures within 12 h. In the presence of serum and FGF-2, cells treated with BPC for 24, 48 and 72 h had no changes in morphology, viability or LDH release compared with the control group. Cell migration in the BPC-treated group was significantly increased compared with the control group. During this process, NO production and ROS level were elevated dramatically, and the levels of HIF-1α and VEGF were increased dependent on the generation of ROS.

Conclusion:

BPC most effectively promoted angiogenesis and migration in HUVECs in the absence of FGF-2 and serum.     

Measurement of the penetration of 56 cosmetic relevant chemicals into and through human skin using a standardized protocol

OECD test guideline 428 compliant protocol using human skin was used to test the penetration of 56 cosmetic-relevant chemicals. The penetration of finite doses (10 μL/cm²) of chemicals was measured over 24 hours. The dermal delivery (DD) (amount in the epidermis, dermis and receptor fluid [RF]) ranged between 0.03 ± 0.02 and 72.61 ± 8.89 μg/cm². The DD of seven chemicals was comparable with in vivo values. The DD was mainly accounted for by the amount in the RF, although there were some exceptions, particularly of low DD chemicals. While there was some variability due to cell outliers and donor variation, the overall reproducibility was very good. As six chemicals had to be applied in 100% ethanol due to low aqueous solubility, we compared the penetration of four chemicals with similar physicochemical properties applied in ethanol and phosphate-buffered saline. Of these, the DD of hydrocortisone was the same in both solvents, while the DD of propylparaben, geraniol and benzophenone was lower in ethanol. Some chemicals displayed an infinite dose kinetic profile; whereas, the cumulative absorption of others into the RF reflected the finite dosing profile, possibly due to chemical volatility, total absorption, chemical precipitation through vehicle evaporation or protein binding (or a combination of these). These investigations provide a substantial and consistent set of skin penetration data that can help improve the understanding of skin penetration, as well as improve the prediction capacity of in silico skin penetration models.     

Growth inhibition and induction of G(1) phase cell cycle arrest in neuroblastoma SH-SY5Y cell by tri-ortho-cresyl phosphate

It has been known that tri-ortho-cresyl phosphate (TOCP) can induce delayed neurotoxicity in humans and sensitive animal species; however, it also has influence on the developing central nervous system or differentiating neuronal cells. In this study, the effects of TOCP on cell proliferation and cell cycle regulation and the mechanisms that contribute to this effect were investigated by using human neuroblastoma SH-SY5Y cell line. Treatment of the cells with TOCP suppressed cell proliferation and reduced cell viability in a dose- and time-dependent manner. Analysis of cell cycle profile indicated that TOCP blocked cell cycle progression by arresting the cell cycle at G(1) phase. The data of determination of cell cycle regulated molecules at mRNA and protein levels showed that TOCP decreased cyclin D1 and increased p21 expression, while did not affect the p53 and p27 levels. Thus, these results indicated that TOCP might induce potential neurodevelopmental toxicity, and a possible mechanism of this toxicity might be the disturbance of cell proliferation by disrupting cell cycle regulatory proteins cyclin D1 and p21 expression.     

para‐Phenylenediamine induces apoptosis through activation of reactive oxygen species‐mediated mitochondrial pathway,and inhibition of the NF‐κB,mTOR, and Wnt pathways in human urothelial cells

para‐Phenylenediamine (PPD) has long been used in two‐thirds of permanent oxidative hair dye formulations. Epidemiological studies and in vivo studies have shown that hair dye is a suspected carcinogen of bladder cancer. However, the toxicity effects of PPD to human bladder remains elusive. In this study, the effects of PPD and its involvement in the apoptosis pathways in human urothelial cells (UROtsa) was investigated. It was demonstrated that PPD decreased cell viability and increased the number of sub‐G₁ hypodiploid cells in UROtsa cells. Cell death due to apoptosis was detected using Annexin V binding assay. Further analysis showed PPD generated reactive oxygen species (ROS), induced mitochondrial dysfunction through the loss of mitochondrial membrane potential and increased caspase‐3 level in UROtsa cells. Western blot analysis of PPD‐treated UROtsa cells showed down‐regulation of phosphorylated proteins from NF‐κB, mTOR, and Wnt pathways. In conclusion, PPD induced apoptosis via activation of ROS‐mediated mitochondrial pathway, and possibly through inhibition of NF‐κB, mTOR, and Wnt pathways. © 2016 Wiley Periodicals, Inc. Environ Toxicol 32: 265–277, 2017.     

The involvement of diadenosine 5',5"'-P1,P4-tetraphosphate in cell cycle arrest and regulation of apoptosis

Diadenosine oligophosphates (Ap(n)A) have been proposed to function as intracellular and extracellular signaling molecules in animal cells. Here, we have examined the cellular and molecular mechanisms underlying the induction of apoptosis by diadenosine 5',5"'-P(1),P(4)-tetraphosphate (Ap(4)A). We have shown a dose-dependent apoptotic response in cells treated with Ap(4)A. Flow cytometric analysis indicated an involvement of Ap(4)A at an early stage of G1/S arrest. No difference in the amount of p21(waf1) was observed in HL60 cells treated with Ap(4)A compared to control cells. The level of retinoblastoma protein (pRb) dropped dramatically when apoptosis was extensive. The cleavage of pRb was abrogated if Ap(4)A-treated cells were incubated with general caspase inhibitor zVAD-fmk. Ap(4)A also induced a profound decrease in the level of the Bcl-2 protein. The lack of effect of Ap(4)A on CDK1 activity indicated that Ap(4)A is not involved in "aberrant mitosis". We suggest that in vivo Ap(4)A may play a significant role in tumor growth suppression by inducing apoptosis.     

Detection and phase I metabolism of the 7‐azaindole‐derived synthetic cannabinoid 5F‐AB‐P7AICA including a preliminary pharmacokinetic evaluation

In June 2018, a 'research chemica'l labeled 'AB‐FUB7AICA' was purchased online and analytically identified as 5F‐AB‐P7AICA, the 7‐azaindole analog of 5F‐AB‐PINACA. Here we present data on structural characterization, suitable urinary consumption markers, and preliminary pharmacokinetic data. Structure characterization was performed by nuclear magnetic resonance spectroscopy, gas chromatography–mass spectrometry, infrared and Raman spectroscopy. Phase I metabolites were generated by applying a pooled human liver microsome assay (pHLM) to confirm the analysis results of authentic urine samples collected after oral self‐administration of 2.5 mg 5F‐AB‐P7AICA. Analyses of pHLM and urine samples were performed by liquid chromatography?time‐of‐flight mass spectrometry and liquid chromatography–tandem mass spectrometry (LC–MS/MS). An LC–MS/MS method for the quantification of 5F‐AB‐P7AICA in serum was validated. Ten phase I metabolites were detected in human urine samples and confirmed in vitro. The main metabolites were formed by hydroxylation, amide hydrolysis, and hydrolytic defluorination, though – in contrast with most other synthetic cannabinoids – the parent compound showed the highest signals in most urine samples. The compound detection window was more than 45 hours in serum. The concentration‐time profile was best explained by a two‐phase pharmacokinetic model. 5F‐AB‐P7AICA was detected in urine samples until 65 hours post ingestion. Monitoring of metabolite M07, hydroxylated at the alkyl chain, next to parent 5F‐AB‐P7AICA, is recommended to confirm the uptake of 5F‐AB‐P7AICA in urinalysis. It seems plausible that the shift of the nitrogen atom from position 2 to 7 (e.g. 5F‐AB‐PINACA to 5F‐AB‐P7AICA) leads to a lower metabolic reactivity, which might be of general interest in medicinal chemistry.     

8-OH-DPAT as a 5-HT7 agonist: phase shifts of the circadian biological clock through increases in cAMP production

Neurons in the suprachiasmatic nucleus (SCN), the site of the endogenous biological clock in mammals, fire spontaneously, peaking in firing rate near ZT6 or at the midpoint of the light phase in a 12:12 light-dark cycle. In rat hypothalamic slices, tissue incubations with drugs can produce a shift in this daily rhythm, the magnitude of which is dependent upon dose and the time of treatment. Previous work with 8-OH-DPAT had noted its ability to produce a phase advance, an earlier occurrence of the peak in neuronal firing, when applied at ZT6. Activation of 5-HT7 receptors was thought to be responsible for the shift, despite the clear preference of 8-OH-DPAT for 5-HT1A sites in terms of receptor binding affinity. In the present work, the actions of 8-OH-DPAT in SCN slices were confirmed and expanded to include additional dose-response and antagonist treatments. By itself, 8-OH-DPAT produced a concentration-dependent phase advance that was sensitive to co-application with 5-HT7 antagonists (ritanserin, mesulergine, SB-269970), but not to 5-HT1A antagonists (WAY-100,635, UH-301). Assignment of the receptor mechanisms for the antagonists employed was accomplished in experiments measuring binding affinities and the generation of cAMP, the latter monitored in a HEK-293 cell line expressing the r5-HT7 receptor and in tissue derived from rat SCN. The results indicate that the increases observed in cAMP levels are small but appear to be sufficient to produce a pharmacological resetting of the clock pacemaker. By aiding in the identification of the 5-HT receptor subtype responsible for the observed phase shifts and cAMP changes, 8-OH-DPAT represents an important pharmacological tool for 5-HT7 receptor activation, essentially broadening its role as the prototypical 5-HT1A agonist to one combining these two receptor activities.     

Liraglutide attenuates gefitinib-induced cardiotoxicity and promotes cardioprotection through the regulation of MAPK/NF-κB signaling pathways

Gefitinib is an effective treatment for patients with locally advanced non-small cell lung cancer. However, it is associated with cardiotoxicity that can limit its clinical use. Liraglutide, a glucagon-like peptide 1 receptor agonist, showed potent cardioprotective effects with the mechanism is yet to be elucidated. Therefore, this study aimed to determine the efficiency of liraglutide in protecting the heart from damage induced by gefitinib. Adult male Wistar rats were randomly divided into control group, liraglutide group (200 µg/kg by i.p. injection), gefitinib group (30 mg/kg orally) and liraglutide plus gefitinib group. After 28 days, blood and tissue samples were collected for histopathological, biochemical, gene and protein analysis. We demonstrated that gefitinib treatment (30 mg/kg) resulted in cardiac damage as evidenced by histopathological studies. Furthermore, serum Creatine kinase-MB (CK-MB), N-terminal pro B-type natriuretic peptide (NT-proBNP) and cardiac Troponin-I (cTnI) were markedly elevated in gefitinib group. Pretreatment with liraglutide (200 µg/kg), however, restored the elevation in serum markers and diminished gefitinib-induced cardiac damage. Moreover, liraglutide improved the gene and protein levels of anti-oxidant (superoxide dismutase) and decreased the oxidative stress marker (NF-κB). Mechanistically, liraglutide offered protection through upregulation of the survival kinases (ERK1/2 and Akt) and downregulation of stress-activated kinases (JNK and P38). In this study, we provide evidence that liraglutide protects the heart from gefitinib-induced cardiac damage through its anti-oxidant property and through the activation of survival kinases.     

Concentration‐dependent induction of reactive oxygen species,cell cycle arrest and apoptosis in human liver cells after nickel nanoparticles exposure

Due to advent of nanotechnology, nickel nanoparticles (Ni NPs) are increasingly recognized for their utility in various applications including catalysts, sensors and electronics. However, the environmental and human health effects of Ni NPs have not been fully investigated. In this study, we examined toxic effects of Ni NPs in human liver (HepG2) cells. Ni NPs were prepared and characterized by X‐ray diffraction, transmission electron microscopy and dynamic light scattering. We observed that Ni NPs (size, ～28 nm; concentration range, 25–100 μg/mL) induced cytotoxicity in HepG2 cells and degree of induction was concentration‐dependent. Ni NPs were also found to induce oxidative stress in dose‐dependent manner evident by induction of reactive oxygen species and depletion of glutathione. Cell cycle analysis of cells treated with Ni NPs exhibited significant increase of apoptotic cell population in subG1 phase. Ni NPs also induced caspase‐3 enzyme activity and apoptotic DNA fragmentation. Upregulation of cell cycle checkpoint gene p53 and bax/bcl‐2 ratio with a concomitant loss in mitochondrial membrane potential suggested that Ni NPs induced apoptosis in HepG2 cells was mediated through mitochondrial pathway. This study warrants that applications of Ni NPs should be carefully assessed as to their toxicity to human health. © 2013 Wiley Periodicals, Inc. Environ Toxicol 30: 137–148, 2015.     

Wogonin potentiates the antitumor effects of low dose 5-fluorouracil against gastric cancer through induction of apoptosis by down-regulation of NF-kappaB and regulation of its metabolism

Traditional Chinese medicines have been recognized as a new source of anticancer drugs or chemotherapy adjuvant to enhance the efficacy of chemotherapy and to ameliorate the side effects. Wogonin (WOG) has a potential for therapeutic use in the treatment of antitumor and chemoprophylaxis. 5-Fluorouracil (5-FU) is a key systemic chemotherapy drug and widely use in the treatment of solid tumors. In this study, we found that combination of WOG and 5-FU inhibited the viability of MGC-803 cells in a concentration-dependent manner and exhibited a synergistic anticancer effect (CI < 1) when 5-FU was used at relatively low concentrations. The pro-apoptotic activity of two-drug combination was much stronger than single. Furthermore, WOG could decrease the mRNA levels of dihydropyrimidine dehydrogenase (DPD), the metabolic enzymes of 5-FU. WOG could inhibit the NF-κB nuclear translocation and I-κB phosphorylation. Moreover, combined treatment caused significantly growth inhibition of human tumor xenografts. In addition, WOG markedly enhanced the antitumor activity of low dose 5-FU (i.p. 10 mg/kg/day), however there is no toxicity and influence on diet consumption in experimental animals. Taken together, our data's showed that WOG increased 5-FU retention for a prolonged catabolism by modulating 5-FU metabolic enzymes and sensitized the MGC-803 cells to 5-FU induced apoptosis by inhibiting the NF-κB nuclear translocation. The anti-gastric cancer effect of two-drug combination was much stronger than that of WOG or 5-FU alone. These results may be relevant to design new clinical therapeutic strategies against gastric cancer in future.     

微小 RNA-425-5p靶向调控 PTEN表达促进宫颈癌海拉细胞增殖、侵袭和迁移的实验研究

目的 探讨微小RNA(miR)-425-5p对宫颈癌海拉细胞增殖、侵袭和迁移的影响及其机制.方法 2018年10月至2019年10月,利用Lipofectamine TM 2000将miR-425-5p模拟物(mimics)、抑制剂(inhibitor)及其阴性对照转染至海拉细胞中,采用实时荧光定量PCR检测细胞中miR-425-5p的表达情况,MTT法检测细胞增殖活力,平板克隆形成实验检测细胞克隆形成能力,Transwell小室实验检测细胞侵袭和迁移,蛋白质印迹法(Western blotting)检测细胞中磷酸酶及张力蛋白同源基因(PTEN)蛋白的表达.采用生物信息学软件预测、双荧光素酶报告基因实验验证miR-425-5p和PTEN的靶向关系.将PTEN过表达质粒pcDNA3.1-PTEN和PTEN干扰序列siRNA-PTEN转染至海拉细胞后,观察PTEN对海拉细胞增殖活力、克隆形成能力、侵袭能力和迁移能力的影响.结果 与各自阴性对照比较,miR-425-5p过表达可促进海拉细胞增殖[(142.25±11.32)％比(100.00±6.67)％]、克隆形成、侵袭[(133.28±9.86)个比(77.46±5.32)个]和迁移[(187.56±15.12)个比(115.35±10.26)个]并下调PTEN蛋白表达,而miR-425-5p低表达则抑制海拉细胞增殖[(58.38±3.45)比(100.00±5.74)]、克隆形成、侵袭[(43.32±3.62)个比(75.65±6.15)个]和迁移[(62.28±4.05)个比(109.72±9.84)个]并上调PTEN蛋白表达(P<0.05).双荧光素酶报告基因实验证实,PTEN是miR-425-5p的靶基因;PTEN过表达可促进海拉细胞增殖[(66.52±4.36)％比(100.00±6.18)％]、克隆形成、侵袭[(46.23±3.13)个比(71.65±5.24)个]和迁移[(62.65±4.26)个比(108.42±8.57)个],而PTEN低表达则抑制海拉细胞增殖[(136.52±9.85)个比(100.00±7.05)个]、克隆形成、侵袭[(110.27±9.85)个比(70.52±6.18)个]和迁移[(168.78±16.96)个比(113.26±10.13)个].结论 miR-425-5p可通过靶向调控PTEN表达促进宫颈癌海拉细胞增殖、侵袭和迁移.     

Quercetin promotes gastrointestinal motility and mucin secretion in loperamide-induced constipation of SD rats through regulation of the mAChRs downstream signal

Context: Quercetin (QCT) has been known as a potential therapeutic strategy for gastrointestinal diseases because it contributes to the stabilization of mast cells, the prevention of histamine release and modulation of CaCC chloride channel.

Objective: We investigated the laxative effect and action mechanism of QCT in Lop-induced constipation model.

Materials and methods: Constipation of SD rats was induced by subcutaneous injection of loperamide (Lop) (4 mg/kg weight) in 0.5% Tween 20 twice a day for three days. After 24?h, the constipation group was further treated with 1× PBS (Lop?+?Vehicle treated group), 10?mg/kg of QCT (Lop?+?LQCT treated group), 20?mg/kg of QCT (Lop?+?MQCT treated group) or 40?mg/kg QCT (Lop?+?HQCT treated group) at once. At 24?h after QCT treatment, the constipation phenotypes were measured and the transverse colon was collected from SD rats.

Results: The gastrointestinal motility, the number of stools and histological structures were significantly recovered in Lop?+?QCT treated group compared with the Lop?+?Vehicle treated group. Also, above activity of epithelial cells and smooth muscle cells were regulated by the mRNA expression of the muscarinic acetylcholine receptors M2 and M3 (mAChR M2 and M3) and some mediators of their downstream signalling pathway. Finally, laxative effects of QCT on mAChR signalling pathway were significantly inhibited by the treatment of mAChR antagonist in primary smooth muscle of rat intestine cells (pRISMCs).

Conclusions: This study provides the first strong evidence that QCT can be considered an important candidate for improving chronic constipation induced by Lop treatment in animal models.     

川芎嗪联合mTOR抑制剂调控卵巢癌SKOV-3细胞增殖、侵袭迁移的实验研究

目的 研究川芎嗪联合mTOR抑制剂对卵巢癌SKOV-3细胞增殖、侵袭迁移的作用.方法 分别设置对照组、40μmol/L川芎嗪组、5μmol/L雷帕霉素组、40μmol/L川芎嗪联合5μmol/L雷帕霉素组,各组SKOV-3细胞给药后继续培养48 h后,MTT法考察各组SKOV-3细胞的增殖抑制率,Transwell实验...     

Geniposide promotes the proliferation and differentiation of MC3T3-E1 and ATDC5 cells by regulation of microRNA-214

The research plans to make sure how Geniposide (GEN) functions in osteoblast proliferation and differentiation. The MC3T3-E1 and ATDC5 cells were treated with the GEN, XAV-939 and/or transfected with microRNA (miR)-214 mimic or corresponding control. Cell viability was detected with the CCK-8. The CyclinD1, Runx2, Osx, Ocn, Wnt3a and β-catenin were individually quantified via western blot. The cell cycle was tested by cell cycle analysis assay. The ALP activity was tested by ALP assay. qRT-PCR was used to examine the miR-214 expression level. The cell viability and the expressions of the CyclinD1, Runx2, Osx, Ocn Wnt3a and β-catenin, as well as the ALP activity were individually and significantly promoted by the GEN. Besides, miR-214 was down-regulated by the GEN. The XAV-939 or the miR-214 mimic destroyed the promotional effect of GEN on these elements above. In conclusion, GEN induced the proliferation and differentiation of the MC3T3-E1 and ATDC5 cells by targeting the miR-214 through Wnt/β-catenin activation.     

Crude extract of Rheum palmatum L. Induces cell cycle arrest S phase and apoptosis through mitochondrial‐dependent pathways in U‐2 OS human osteosarcoma cells

Cancer is the second cause of death in children. Osteosarcoma is the most common primary malignancy of solid bone cancer primarily affecting adolescents and young adults. In the Chinese population, the crude extract of Rheum palmatum L. (CERP) has been used for treating different diseases, including SARS, rheumatoid arthritis, coxsackievirus B3, and human colon cancer cell, pancreatic cancer. There are no reports on CERP and human osteosarcoma cells. The present study examined effects of CERP on cytotoxicity including cell cycle distribution and cell death (apoptosis) in U‐2 OS human osteosarcoma cells. CERP significantly induced S phase arrest in U‐2 OS cells in a dose‐dependent. CERP produced DNA damage and DNA condensation. Other effects of CERP were stimulation of ROS and Ca²⁺, mitochondria impairment, and activation of caspase‐3, ?8, and ?9. CERP increased the levels of Bax, Bak, Bad, cyclin B, Fas, PARP, GRP78, GADD153, AIF, Endo G, Calpain‐2, p21, and p27, but decreased the levels of Bcl‐2, BCL‐X, XIAP, Akt, CDC25A, CDK2, Cyclin A, and Cyclin E of U‐2 OS cells. It was also observed that CERP promoted the expression of AIF, Endo G, GADD153, and cytochrome c. These results indicate that CERP has anticancer effects in vitro and provide the foundation for in vivo studies of animal models of osteosarcoma. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 957–969, 2016.     

Cytotoxic, mutagenic and genotoxic effects of new anti-T. cruzi 5-phenylethenylbenzofuroxans. Contribution of phase I metabolites on the mutagenicity induction

5-Phenylethenylbenzofuroxans have displayed in vitro and in vivo activity against Trypanosoma cruzi, the etiologic agent of American Trypanosomiasis. On the basis of benzofuroxans pre-clinical studies we evaluated the potential of six 5-phenylethenyl derivatives to induce cytotoxicity, mutagenicity and genotoxicity using different in vitro models. Cytotoxic effects were evaluated using a set of cells, mammal pre-monocytic macrophages, V-79 lung fibroblast from Chinese hamster, and colorectal adenocarcinoma Caco-2 cells, in the MTT viability assay. Mutagenicity was tested in the Ames assay using Salmonella typhimurium TA98 strain with and without metabolic activation by S9-rat liver homogenate. The genotoxic potentials were evaluated with the alkaline single cell gel electrophoresis (comet assay) in V-79 cells. In view of the Ames test results we study whether the main mammals’ phase I metabolites, the corresponding o-nitroanilines, are involved in the mechanism of mutagenicity. These metabolites are produced by NADPH-dependent enzymes in cytosol and by xanthine oxidase and cytochrome P450 in microsomes from rat liver. Among them, the electronic property of phenyl substituent seems to be responsible for this effect. It could be pointed out that the equimolecular mixture of compounds 1 and 2 (5E- and 5Z-(2-phenylethenyl)benzofuroxan, respectively) could be used in further clinical studies as anti-T. cruzi drug.